Increased in vivo inhibition of gene expression by combining RNA interference and U1 inhibition
Increased in vivo inhibition of gene expression by combining RNA interference and U1 inhibition
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DOI：
10.1093/nar/gkr956
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发表时间：
2012-01-01
影响因子：
14.9
通讯作者：
Fortes, Puri
中科院分区：
文献类型：
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作者：
Blazquez, Lorea;Jovanna Gonzalez-Rojas, Sandra;Fortes, Puri
抑制基因表达可通过RNA干扰(RNAi)或U1小核RNA-SnRNA干扰(U1i)实现。U1i是基于U1抑制物(U1in)，U1SnRNA分子经修饰后抑制多聚腺苷酸化的一种靶标前-mRNA。在培养中，我们已经表明，RNAi和U1i的组合比单独使用这两种技术对报告基因或内源基因的抑制更强。我们现在已经使用这些技术来抑制小鼠的基因表达。我们发现U1ins可以在体内诱导对靶基因表达的强烈抑制。此外，U1i和RNAi的结合也会在小鼠中产生协同抑制。这表现为对乙肝病毒(HBV)序列或内源性Notch1的抑制。令人惊讶的是，U1in和RNAi介体结合获得的抑制作用比两个U1in或两个RNAi介体结合获得的抑制作用要高。我们的结果表明，RNAi和U1i通过未知的机制相互作用，导致协同抑制。毒性和特异性分析表明，U1I抑制剂的表达是安全的。因此，我们认为，RNAi和U1i的结合将是阻断体内破坏性内源性基因、乙肝病毒和其他感染源的良好选择。
Inhibition of gene expression can be achieved with RNA interference (RNAi) or U1 small nuclear RNA-snRNA-interference (U1i). U1i is based on U1 inhibitors (U1in), U1 snRNA molecules modified to inhibit polyadenylation of a target pre-mRNA. In culture, we have shown that the combination of RNAi and U1i results in stronger inhibition of reporter or endogenous genes than that obtained using either of the techniques alone. We have now used these techniques to inhibit gene expression in mice. We show that U1ins can induce strong inhibition of the expression of target genes in vivo. Furthermore, combining U1i and RNAi results in synergistic inhibitions also in mice. This is shown for the inhibition of hepatitis B virus (HBV) sequences or endogenous Notch1. Surprisingly, inhibition obtained by combining a U1in and a RNAi mediator is higher than that obtained by combining two U1ins or two RNAi mediators. Our results suggest that RNAi and U1i cooperate by unknown mechanisms to result in synergistic inhibitions. Analysis of toxicity and specificity indicates that expression of U1i inhibitors is safe. Therefore, we believe that the combination of RNAi and U1i will be a good option to block damaging endogenous genes, HBV and other infectious agents in vivo.